Therapeutic peptides and uses thereof

Abstract

Methods for ameliorating neurodegenerative disorders and methods for ameliorating inflammatory disorders comprising administering to said subject an effective amount of a polypeptide comprising, in order from N-terminus to C-terminus, R-X.sub.1-X.sub.2-X.sub.3-X.sub.4-X.sub.5-R wherein: R is optionally 1-5 additional ?-amino acids of either L-configuration or D configuration; X.sub.1 is any amino acid of either L-configuration or D-configuration; X.sub.2 is any amino acid of either L-configuration or D-configuration; X.sub.3 is Asp or Glu of either L-configuration or D-configuration; X.sub.4 is any amino acid of either L-configuration or D-configuration; X.sub.5 is any amino acid of either L-configuration or D-configuration; and R is optionally 1-5 additional ?-amino acids of either L-configuration or D-configuration, with the proviso that at least three of X.sub.1, X.sub.2, X.sub.3, X.sub.4, and X.sub.5 are of the D-configuration, wherein the N-terminus is optionally modified by acetylation, and wherein the C-terminus is optionally modified by amidation and/or methylation.

Claims

1. A method for ameliorating amyotrophic lateral sclerosis (ALS) in a subject suffering from ALS comprising administering to said subject an effective amount of a polypeptide selected from the group consisting of: a polypeptide comprising amino acid sequence D-Tyr-D-Val-D-Asp-D-Lys-D-Arg (SEQ ID NO:1), a polypeptide comprising amino acid sequence D-Tyr-D-Val-D-Asp-D-Lys-D-Arg-NH.sub.2 (SEQ ID NO:2), a polypeptide comprising amino acid sequence Ac-D-Tyr-D-Val-D-Asp-D-Lys-D-Arg-NH.sub.2 (SEQ ID NO:3), a polypeptide comprising amino acid sequence D-Tyr-D-Val-D-Asp-D-Lys-D-Arg-NHMe (SEQ ID NO:4), a polypeptide comprising amino acid sequence Ac-D-Tyr-D-Val-D-Asp-D-Lys-D-Arg-NHMe (SEQ ID NO:5), and a polypeptide comprising amino acid sequence D-Tyr-D-Val-D-Asp-D-Lys-D-Asp-NH.sub.2 (SEQ ID NO:7).

2. The method according to claim 1, wherein the polypeptide consists of an amino acid sequence set forth in SEQ ID NO: 1.

3. The method according to claim 1, wherein said polypeptide is administered intrathecally, intravenously, orally, and/or subcutaneously.

4. The method according to claim 1, wherein said polypeptide is administered in an amount ranging from 1 mg/kg of said subject to 50 mg/kg of said subject.

5. The method according to claim 1, wherein said polypeptide is administered in an amount ranging from 1 mg/kg of said subject to 10 mg/kg of said subject.

6. The method according to claim 1, wherein said polypeptide is administered in an amount ranging from 1 mg/kg of said subject to 1 mg/kg of said subject.

7. The method according to claim 1, wherein said polypeptide is lyophilized and reconstituted with an appropriate amount of diluent selected from the group consisting of distilled water and/or sodium chloride.

8. The method according to claim 2, wherein said polypeptide is administered in an amount of 0.04 mg/kg of said subject given every 48 hours.

9. The method according to claim 2, wherein said polypeptide is administered in a total amount of 2 mg/kg of said subject every other day.

10. The method according to claim 2, wherein said polypeptide is administered subcutaneously in an amount of 0.04 mg/kg of said subject given every 48 hours.

11. The method according to claim 1, wherein the polypeptide consists of the amino acid D-Tyr-D-Val-D-Asp-D-Lys-D-Arg (SEQ ID NO:1), D-Tyr-D-Val-D-Asp-D-Lys-D-Arg-NH.sub.2 (SEQ ID NO:2), Ac-D-Tyr-D-Val-D-Asp-D-Lys-D-Arg-NH.sub.2 (SEQ ID NO:3), D-Tyr-D-Val-D-Asp-D-Lys-D-Arg-NHMe (SEQ ID NO:4), Ac-D-Tyr-D-Val-D-Asp-D-Lys-D-Arg-NHMe (SEQ ID NO:5), or D-Tyr-D-Val-D-Asp-D-Lys-D-Asp-NH.sub.2 (SEQ ID NO:7).

12. A method for ameliorating chronic renal disease, renal disease, and/or end-stage renal disease in a subject suffering therefrom comprising administering to said subject an effective amount of a polypeptide selected from the group consisting of: a polypeptide comprising amino acid sequence D-Tyr-D-Val-D-Asp-D-Lys-D-Arg (SEQ ID NO:1), a polypeptide comprising amino acid sequence D-Tyr-D-Val-D-Asp-D-Lys-D-Arg-NH.sub.2 (SEQ ID NO:2), a polypeptide comprising amino acid sequence Ac-D-Tyr-D-Val-D-Asp-D-Lys-D-Arg-NH.sub.2 (SEQ ID NO:3), a polypeptide comprising amino acid sequence D-Tyr-D-Val-D-Asp-D-Lys-D-Arg-NHMe (SEQ ID NO:4), a polypeptide comprising amino acid sequence Ac-D-Tyr-D-Val-D-Asp-D-Lys-D-Arg-NHMe (SEQ ID NO:5), and a polypeptide comprising amino acid sequence D-Tyr-D-Val-D-Asp-D-Lys-D-Asp-NH.sub.2 (SEQ ID NO:7).

13. The method according to claim 12, wherein the polypeptide consists of an amino acid sequence set forth in SEQ ID NO: 1.

14. The method according to claim 12, wherein said polypeptide is administered intrathecally, intravenously, orally, and/or subcutaneously.

15. The method according to claim 12, wherein said polypeptide is administered in an amount ranging from 1 mg/kg of said subject to 50 mg/kg of said subject.

16. The method according to claim 12, wherein said polypeptide is administered in an amount ranging from 1 mg/kg of said subject to 10 mg/kg of said subject.

17. The method according to claim 12, wherein said polypeptide is administered in an amount ranging from 1 mg/kg of said subject to 1 mg/kg of said subject.

18. The method according to claim 12, wherein said polypeptide is lyophilized and reconstituted with an appropriate amount of diluent selected from the group consisting of distilled water and/or sodium chloride.

19. The method according to claim 13, wherein said polypeptide is administered in an amount of 0.04 mg/kg of said subject given every 48 hours.

20. The method according to claim 13, wherein said polypeptide is administered in a total amount of 2 mg/kg of said subject every other day.

21. The method according to claim 13, wherein said polypeptide is administered subcutaneously in an amount of 0.04 mg/kg of said subject given every 48 hours.

22. The method according to claim 12, wherein the polypeptide consists of the amino acid sequence D-Tyr-D-Val-D-Asp-D-Lys-D-Arg (SEQ ID NO:1), D-Tyr-D-Val-D-Asp-D-Lys-D-Arg-NH.sub.2 (SEQ ID NO:2), Ac-D-Tyr-D-Val-D-Asp-D-Lys-D-Arg-NH.sub.2 (SEQ ID NO:3), D-Tyr-D-Val-D-Asp-D-Lys-D-Arg-NHMe (SEQ ID NO:4), Ac-D-Tyr-D-Val-D-Asp-D-Lys-D-Arg-NHMe (SEQ ID NO:5), or D-Tyr-D-Val-D-Asp-D-Lys-D-Asp-NH.sub.2 (SEQ ID NO:7).

Description

BRIEF DESCRIPTION OF THE DRAWINGS

(1) The foregoing will be apparent from the following more particular description of example embodiments of the invention, as illustrated in the accompanying drawings.

(2) FIG. 1 shows that TVALA? (SEQ ID NO:1) given at 10 mg/kg orally had smaller phrenic axons than vehicle treated mice.

(3) FIG. 2 shows that TVALA? (SEQ ID NO:1) given at 10 mg/kg orally had smaller phrenic axons than vehicle treated mice.

(4) FIG. 3 shows that TVALA? (SEQ ID NO:1) given at 3 mg/kg orally had slightly larger femoral motor axons than vehicle treated mice.

(5) FIG. 4 shows that TVALA? (SEQ ID NO:1) given at 3 mg/kg orally had more significant effects than 10 mg/kg orally on the neuroinflammatory marker GFAP demonstrating the biphasic action of a neurotropic molecule.

(6) FIG. 5 shows that TVALA? (SEQ ID NO:1) decreased extracellular ATP, eATP (circle) in in vitro cultured adipose stem cells (ASC) from an ALS patient. The horizontal line indicates the dysfunctional levels in untreated cells, the box indicates the desired concentration of TVALA? that rescues the mitochondria in ALS-ASCs. The X axis is TVALA? concentration and the Y axis is % eATP.

(7) FIGS. 6A-6C show that the peptide TVALA? (SEQ ID NO:1) used to pretreat A-375 melanoma cells did not rescue the cells against Erastin-induced cell death. FIG. 6D is the legend for FIGS. 6A-6C.

(8) FIGS. 7A-7C show that the peptide TVALA? (SEQ ID NO:1), pretreatment followed by incubation with peptide followed by Erastin, the cells were protected against Erastin-induced cell death. FIG. 7D is the legend for FIGS. 7A-7C.

(9) FIG. 8 shows 32 growth factors and cytokines in optimized culture media from adipose stem cells (ASCs) from a healthy donor.

(10) FIG. 9 shows growth factors and cytokines in optimized culture media from adipose stem cells (ASCs) from an ALS patient.

DETAILED DESCRIPTION OF THE INVENTION

(11) The inventor has determined that polypeptides, including H-(D-Tyr)-(D-Val)-(D-Asp)-(D-Lys)-(D-Arg)-OH (SEQ ID NO:1), are valuable tools at ameliorating neurodegenerative and inflammatory disorders, including ALS, AD, PD, neurodegenerative disorders, inflammatory disorders (including ankylosing spondylitis, Crohn's disease, hidradenitis suppurativa, juvenile idiopathic arthritis, non-radiographic axial spondyloarthritis, non-infectious uveitis, plaque psoriasis, psoriasis, psoriatic arthritis, rheumatoid arthritis, and ulcerative colitis), and dysregulated mitochondrial energy metabolism.

(12) The polypeptides comprise, in order from N-terminus to C-terminus,
R-X.sub.1-X.sub.2-X.sub.3-X.sub.4-X.sub.5-R
wherein: R is optionally 1-5 additional ?-amino acids of either L-configuration or D-configuration; X.sub.1 is any amino acid of either L-configuration or D-configuration; X.sub.2 is any amino acid of either L-configuration or D-configuration; X.sub.3 is Asp or Glu of either L-configuration or D-configuration; X.sub.4 is any amino acid of either L-configuration or D-configuration; X.sub.5 is any amino acid of either L-configuration or D-configuration; and R is optionally 1-5 additional ?-amino acids of either L-configuration or D-configuration,
with the proviso that at least three of X.sub.1, X.sub.2, X.sub.3, X.sub.4, and X.sub.5 are of the D-configuration,
wherein the N-terminus is optionally modified by acetylation, and
wherein the C-terminus is optionally modified by amidation and/or methylation.

(13) The Endoqenous Immune Modulator Thymopoetin and Thymopentin

(14) The thymic hormone thymopoietin is an endogenous anti-inflammatory molecule with multiple actions in multiple tissues. The active part of the molecule is a pentapeptide, represented by the synthetic molecule thymopentin, that binds thymopoietin receptors. The central amino acid of the active pentapeptide and determines the bioactivity. See V. K. Singh et al., Thymopentin and splenopentin as immunomodulators. Curent status, 17(3) IMMUNOL. RES. 345-368 (1998).

(15) Thymopoietin suppresses NF-kB activation, inhibits microglial activation by reducing secretion of inflammatory mediators, increases production of new oligodendrocytes generated from oligodendrocyte progenitor cells and assists their differentiation into mature myelinating oligodendrocytes. See C. Tuthill et al., Thymosin Apha 1 A Peptide Immune Modulator with a Broad Range of Clinical Applications, 3(4) JOURNAL OF CLINICAL AND EXPERIMENTAL PHARMACOLOGY 1-17 (2013). The pro-molecule thymalfasin (Zadaxin?) is metabolized to the active pentapeptide thymopentin. See M. Severa et. al., Thymosins in multiple sclerosis and its experimental models: moving from basic to clinical application, 27 MULTIPLE SCLEROSIS AND RELATED DISORDERS 52-60 (2019). Thymopentin, Arg-Lys-Asp-Val-Tyr, that corresponds to positions 32-36 of thymopoietin, where it induces regulatory T cells and inhibits activated B cell differentiation and is a neurotropic factor.

(16) TABLE-US-00001 TABLE 1 Comparison of three well-studied thymopoietin receptor molecules Thymopoietin Thymalfasin Thymopentin 49 amino acids 28 amino acids 5 amino acids suppresses NF-kB Treats hepatitis Inhibits B cell activation B and C differentiation inhibits microglial Anti-inflammatory Induces T regulatory activation cells Reduces inflammatory Rescues stressed Reduces inflammatory mediators mitochondria mediators Increases Metabolized to Neurotropic factor oligodendrocytes thymopentin Assists differentiation Treats chemotherapy The active part of of oligodendrocytes induced thymopoietin immunosuppression See C. Tuthill et al., Thymosin Apha 1A Peptide Immune Modulator with a Broad Range of Clinical Applications, 3(4) JOURNAL OF CLINICAL AND EXPERIMENTAL PHARMACOLOGY 1-17 (2013)

(17) Thymopentin is the minimal sequence that reproduces the biological activities of thymopoietin and is an immunoregulatory molecule. Products containing thymopentin include Timunox?, Mepentil? and Sintomodulina? which are used for primary immunodeficiencies, secondary immunodeficiencies, in complications of cancer patients after chemotherapy, and for the stimulation of immune responses. Thymopentin is a soluble peptide hormone with pleiotropic effects. Both in vitro and in vivo studies have been conducted to show the multiple roles of thymopentin. See V. K. Singh et al., Thymopentin and splenopentin as immunomodulators. Current status, 17(3) IMMUNOL. RES. 345-368 (1998). Thymopentin demonstrated a dose dependent stimulation or inhibition of mitogen induced IgG, T cell proliferation, and enhanced synthesis of IL-2 in vivo. See J. Duchateau et al., Immunomodulation with thymopentin: in vitro studies, 6(1) MED. ONCOL. TUMOR PHARMACOTHER. 19-23 (1989); J. Duchateau et al., In vitro influence of thymopentin on proliferative responses and phytohemagglutinin-induced interleukin 2 production in normal human lymphocyte cultures, 4 SURVEY OF IMMUNOLOGIC RESEARCH 116-124 (1985). Additional in vivo mitogen induced responses include IFN production and enhanced NK cell activity in mice cells. See W. Diezel et al., Induction and augmentation of mitogen-induced immune interferon production in human lymphocytes by a synthetic thymopoietin pentapeptide, 43(6) BIOMED BIOCHIM ACTA. K9-K12 (1984); C. Hu et al., In vivo enhancement of NK-cell activity by thymopentin, 12(2) INTERNATIONAL JOURNAL OF IMMUNOPHARMACOLOGY 193-197 (1990).

(18) Immunomodulatory actions from in vivo studies demonstrated thymopentin blocked neurotransmission and restored helper T cell activity. See M. C. Weksler et al., Immunological studies of aging. IV. The contribution of thymic involution to the immune deficiencies of aging mice and reversal with thympoietin32-36, 148(4) J. EXP. MED. 996-1006 (1978).

(19) In renal dialysis patients thymopentin reduced IL-6, IL-8, TNF and CRP indicating that treatment notably reduced the inflammatory responses in the body and inhibit an acute phase response induced by end-stage renal disease. See Q. Zou et al., The effect of thymopentin on immune function and inflammatory levels in end-stage renal disease patients with maintenance hemodialysis, 14(1) AM. J. TRANSL. RES. 414-420 (2022). In the same study, thymopentin improved the state of oxidative stress by increasing the SOD, a major antioxidant enzyme that inhibits the accumulation of lipid peroxides.

(20) Multiple studies on the safety of thymopentin found it well tolerated even when administered concomitantly with a long list of drugs given for other reasons. The overall consensus is that thymopentin is a safe compound. See N. Friedmann, Thymopentin: Safety overview, 4 SURVEY OF IMMUNOLOGIC RESEARCH 139-148 (1985).

(21) The Utility of ALS Mouse Models to Test Thymalfasin and Reactive Molecules

(22) The transgenic mouse SOD1.sup.G93A is a toxic gain of function model that overexpresses the human SOD1 dismutase gene. The mice develop adult-onset neurodegeneration of spinal motor neurons and progressive motor deficits leading to paralysis. First reported in 1994, the G93A models are a cornerstone of preclinical ALS research despite the inability to translate efficacy in studies to effectiveness in humans. The model is useful to evaluate peripheral neuropathy using the femoral nerve. Ubiquitin in the brain and spinal cord are drugable targets and in this model ubiquitin ratio indicate impaired autophagy, not neuroinflammation. See C. Cheroni et al., Accumulation of human SOD1 and ubiquitinated deposits in the spinal cord of SOD1G93A mice during motor neuron disease progression correlates with a decrease of proteasome, 18(3) NEUROBIOL DIS. 509-522 (2005); B. Gong et al., The Ubiquitin-Proteasome System: Potential Therapeutic Targets for Alzheimer's Disease and Spinal Cord Injury, 9(Article 4) FRONTIERS IN MOLECULAR NEUROSCIENCE 1-16 (2016).

(23) TAR DNA binding protein plays a crucial role in a growing set of neurodegenerative diseases including ALS. Supporting a central role of TAR DNA Binding Protein 43 in ALS, this protein links both familial and sporadic forms of ALS as mutations that are causative for disease and cytoplasmic aggregates are a hallmark of nearly all cases, regardless of TDP43 mutational status. See T. R. Suk et al., The role of TDP-43 mislocalization in amytrophic lateral sclerosis, 15:45 MOLECULAR NEURODEGENERATION 1-16 (2020). The forgoing supports investigating treatments affecting inflammation in the TDP43 mouse model. Astrocytes represent 30-40% of the cells in the CNS, form an integral part of the blood-brain barrier and establish numerous interactions with other cells in the nervous system including neurons. Astrocytes are central to the normal function of synapses and contribute to axonal metabolic maintenance through the regulation of ion homeostasis. See A. Abdelhak et al., Blood GFAP as an emerging biomarker in brain and spinal cord disorders, 18 NATURE REVIEWS NEUROLOGY 158-172 (2022). Glial fibrillary acidic protein (GFAP) is the signature intermediate filament of astrocytes. The TDP43 transgenic mouse is useful to evaluate inflammation using GFAP levels and Iba1 ratios determined by histopathology.

(24) Evidence that Thymalfasin May Target ALS Related Disease in ALS Mice

(25) In mice, thymalfasin is metabolized to the active molecule thymopentin. Because mice process the pro-molecule thymalfasin into thymopentin as do humans, thymalfasin may illuminate the effects of thymopentin by increasing target engagement afforded by the pro-molecule. Two ALS mouse models, hemizygous B6.Cg-Tg(Prnp-TARDBP*A315T)95Balo/J male mice (JAX stock #010700) the SOD1.sup.G93A model and hemizygous B6SJL-Tg(SOD1*G93A)1Gur/J male mice were dosed with 0.04 mg/kg thymalfasin by subcutaneous (SQ) injection twice a week for 8 weeks as a 1.6 mg lyophilized powder in a 5 ml vial reconstituted with sterile water. Generally accepted markers in the SOD1.sup.G93A and TDP43 models that are associated with neurodegenerative disease include ubiquitin ratio in brain and spinal cord, GFAP, and Iba1. Significant observations in thymalfasin treated ALS mice are shown in Table 2.

(26) TABLE-US-00002 TABLE 2 Significant observations in studies using thymalfasin in two ALS mouse models Significant observations SOD1.sup.G93A model TDP43 model Ubiquitin ratio spinal cord + Ubiquitin ratio brain ++ Brain GFAP ratio ++ Brain Iba1 ++

Definitions

(27) All definitions of substituents set forth below are further applicable to the use of the term in conjunction with another substituent. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

(28) As used herein, the singular forms a, and, and the include plural reference unless the context clearly dictates otherwise. Additionally, the term comprises is intended to include embodiments where the method, apparatus, composition, etc., consists essentially of and/or consists of the listed steps, components, etc. Similarly, the term consists essentially of is intended to include embodiments where the method, apparatus, composition, etc., consists of the listed steps, components, etc.

(29) As used herein, the term about refers to a number that differs from the given number by less than 15%. In other embodiments, the term about indicates that the number differs from the given number by less than 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, or 1%.

(30) As used herein, the term administration is intended to include, but is not limited to, the following delivery methods: topical, oral, sub-lingual, buccal, parenteral, subcutaneous, transdermal, transbuccal, intravascular (e.g., intravenous or intra-arterial), intramuscular, subcutaneous, intranasal, and intra-ocular administration. Administration can be local at a particular anatomical site, such as a site of infection, or systemic.

(31) Arg represents arginine, and is a residue of the amino acid arginine.

(32) Asp represents aspartic acid, and is a residue of the amino acid aspartic acid.

(33) Configuration, as used herein, refers to the orientation of the amino group in a given polypeptide. The term D-configuration refers to a stereoisomer of a particular amino acid whose amino group is on the right side in its Fisher projection. The term L-configuration refers to a stereoisomer of a particular amino acid whose amino group is on the left side in its Fisher projection. If no configuration is listed herein, the L-configuration is to be presumed.

(34) When used herein, a dose range reflected as two numbers means those doses as well as all doses within that range. For example, a dose range from 10 mg-11 mg means 10.0 mg, 10.05 mg, 10.10 mg, 10.15 mg, 10.20 mg, 10.25 mg, 10.30 mg, 10.35 mg, 10.40 mg, 10.45 mg, 10.50 mg, 10.55 mg, 10.60 mg, 10.65 mg, 10.70 mg, 10.75 mg, 10.80 mg, 10.85 mg, 10.90 mg, 10.95 mg, 11.00 mg, as well as any and all amounts therein, such as 10.34 mg, 10.78 mg, etc.

(35) The phrase effective amount means an amount of an agent that (i) treats or prevents the particular disease, condition, or disorder, (ii) attenuates, ameliorates, or eliminates one or more symptoms of the particular disease, condition, or disorder, or (iii) prevents or delays the onset of one or more symptoms of the particular disease, condition, or disorder described herein.

(36) A human subject may be an adult or a child. As used herein, a child refers to a human subject who is between the ages of 1 day to 17 years of age. The term adult refers to a human subject who is 18 years of age or older.

(37) As used herein, a subject is in need of a treatment if such human or non-human animal subject would benefit biologically, medically, or in quality of life from such treatment (preferably, a human).

(38) As used herein, the term inhibit, inhibition, or inhibiting refers to the reduction or suppression of a given condition, symptom, or disorder, or disease, or a significant decrease in the baseline activity of a biological activity or process.

(39) As used herein, the terms subject, patient, and individual are used interchangeably and refer to a human of any age or gender.

(40) As used herein, the term treat, treating, or treatment of any disease or disorder refers in one embodiment, to ameliorating the disease or disorder (i.e., slowing or arresting or reducing the development of the disease or at least one of the clinical symptoms thereof). In another embodiment treat, treating, or treatment refers to alleviating or ameliorating at least one physical parameter including those which may not be discernible by the subject. In yet another embodiment, treat, treating, or treatment refers to modulating the disease or disorder, either physically, (e.g., stabilization of a discernible symptom), physiologically, (e.g., stabilization of a physical parameter), or both. In yet another embodiment, treat, treating, or treatment refers to prophylaxis (preventing or delaying the onset or development or progression of the disease or disorder).

(41) Lys represents lysine, and is a residue of the amino acid lysine.

(42) As used herein, nucleotide sequence or nucleic acid sequence refers to an oligonucleotide sequence or polynucleotide sequence and variants, homologues, fragments, and derivatives thereof. The nucleotide sequence may be of genomic, synthetic, or recombinant origin and may be double-stranded or single-stranded, whether representing the sense or anti-sense strand. As used herein, the term nucleotide sequence includes genomic DNA, cDNA, synthetic DNA, and RNA.

(43) Pharmaceutically-acceptable carrier means non-therapeutic components that are of sufficient purity and quality for use in the formulation of a composition of the invention that, when appropriately administered, typically do not produce an adverse reaction, and that are used as a vehicle for a drug substance.

(44) The phrase pharmaceutically-acceptable indicates that the substance or composition must be compatible chemically and/or toxicologically, with the other ingredients comprising a formulation, and/or the mammal being treated therewith.

(45) Pharmaceutical formulations include pharmaceutically-acceptable and physiologically-acceptable carriers, diluents, or excipients. In this context, the terms pharmaceutically-acceptable and physiologically-acceptable include solvents (aqueous or non-aqueous), solutions, emulsions, dispersion media, coatings, isotonic and absorption promoting or delaying agents, compatible with pharmaceutical administration. Such formulations can be contained in a liquid; emulsion, suspension, syrup, or elixir, or solid form; tablet (coated or uncoated), capsule (hard or soft), powder, granule, crystal, or microbead. Supplementary compounds (e.g., preservatives, antibacterial, antiviral, and antifungal agents) can also be incorporated into the compositions.

(46) As used herein, polypeptide is used interchangeably with the terms amino acid sequence, peptide, and/or protein.

(47) Tyr represents tyrosine, and is a residue of the amino acid tyrosine.

(48) Val represents valine, and is a residue of the amino acid valine.

EXAMPLES

(49) Materials and Methods: The peptides used in the following examples were prepared using standard solid-phase and solution-phase peptide synthesis using techniques well-known in the art. See, e.g., Gregg Fields, Introduction to Peptide Synthesis, CURRENT PROTOCOLS IN PROTEIN SCIENCE (February 2002).

Example 1TVALA? Improves Target Engagement

(50) Thymopentin (TP-5) is a natural, all-L pentapeptide, that is subject to proteolytic cleavage and a short in vivo half-life. The stability in mouse liver microsomes for thymopentin is 23.1 minutes. See Table 3.

(51) TABLE-US-00003 TABLE 3 A comparison of stability of natural thymopentin, TP-5, and TVALA? dosed in mice IV (intravenous), SC (subcutaneous), and PO (orally) Mouse Mouse Plasma Microsomal IV SC PO Stability Stability t?, t?, t?, SC PO SC PO SC PO t? t? mice mice mice % F, % F, Cmax Cmax AUCinf AUCinf Compound (min) (min) (min) (min) (min) mice mice (ng/mL) (ng/ml) hr * ng/mL hr * ng/ml TP-5 23.1 >60 5.58 8.92 7.72 39 36 133 1367 126 1274 TVALA? >120 >60 6.22 11.9 7.36 52 30 674 5291 355 2400

(52) Reversing the orientation of the peptide bonds taken together with inversion of the stereochemistry from the natural L- to the unnatural D- (TVALA?) (SEQ ID NO:1) results in a high degree of topochemical equivalence between the parent peptide and its isomeric replacement. By maintaining the topochemical equivalence the bioactivity of the molecule is maintained. This retro-inverso approach increases the metabolic stability against proteolysis, the stability in mouse liver microsomes has a half-life of over 120 minutes. See Table 3. The increased metabolic stability was also observed in rat and dog microsomes. See Table 4.

(53) TABLE-US-00004 TABLE 4 Stability of TVALA? in mouse, rat, and dog microsomes. % F = % oral bioavailability Microsomal Plasma PO SC IV Stability Stability % F % F t? t? t? B:P (min) (min) (PO) (SC) (hrs) (hrs) (hrs) ratio Mouse >60 >120 30 52 7.4 11.9 6.2 0.03- Rat >60 6.7 79.2 9.6 5.7 6.8 0.06 Dog >60 4.9 3.5

Example 2TVALA? Effects in ALS Animal Models

(54) A 6-week efficacy testing of test article, TVALA?, in daily drinking water, in the SOD1.sup.G93A mouse model of ALS, with neurological scoring, neuroinflammation and axonopathy outcome measures was conducted by orally dosing with 3 mg/kg or 10 mg/kg. In phrenic nerves the axons numbers are not affected at this age, however axons start to atrophy, in femoral nerves, the mice lose axons and the remaining ones are atrophied.

(55) There was very discrete neuroprotection by the 3 mg/kg dose on the phrenic and femoral motor axon sizes (see FIG. 1), the observation was significant from a statistic point of view but the effect is small. The significant results are shown in Table 5. In this study hemizygous mice had fewer femoral axons, wild type mice have mostly larger phrenic nerve axons than untreated hemizygous mice, while 10 mg/kg treated hemizygous mice showed the smallest axons (see FIG. 2). A significant treatment effect with hemizygous mice treated with 3 mg/kg was observed on the femoral motor axon size, treated mice had axons of slightly larger size. See FIG. 3. It is not unusual for neurotropic molecules to have biphasic actions and often with a lower dose being desired as shown in the spinal cord GFAP. See FIG. 4.

(56) These results indicate a low dose may be preferred over a higher dose. Also, brain inflammation was not impacted with oral administration of TVALA? in this study, data not shown. The expected impact of the oral drug on the central nervous system was 5% based on in vivo testing (see Table 4). These results indicate the preferred administration is by sub-cutaneous injection to increase the bioavailability of the drug.

(57) TABLE-US-00005 TABLE 5 Significant results in TVALA? treated SOD1.sup.G93A SOD1.sup.G93A model SOD1.sup.G93A model Observations 3 mg/kg TVALA? 10 mg/kg TVALA? Phrenic nerve cross- Smaller axons than section area control (FIG. 1) Femoral motor Slightly larger than axon size controls (FIG. 3) Spinal cord GFAP Mild sign, better than 3 mg/kg

Example 3Mitogen-Induced Blastogenic Response Assay of Peripheral Blood Mononuclear Cells (PBMCs) Assess Innate Cellular Immunity

(58) The influence of proinflammatory cytokines on the immunoregulatory function of immune cells from healthy donors may be assessed using mitogen-stimulated PBMC responses, which can be measured by determining the effect of test compounds on cytokine levels after stimulation by Concanavilin A (ConA). See Arneth, CURRENT PROTOCOLS IN CYTOMETRY 6.28 supplement 51 (2010); A. Erhardt et al., Immunology and Liver Disease Karger (2010); W. Ge et al., INTERNATIONAL IMMUNOPHARMACOLOGY 85 (2020); S. Malchow et al., BICHIMICA ET BIOPHYSICA ACTA 1812:290-301 (2011); A. Thomazelli et al., IMMUNOPHARMACOLOGY AND IMMUNOTOXICOLOGY, 40:5:387-392 (2018); N. Umeda et al., LEUKOCYTE BIOLOGY 791-801 (2019); H. Wang et al., WORLD JOURNAL OF GASTROENTEROLOGY 119-125 (2012).

(59) Normal human PBMCs were stimulated with 5 ?g/mL ConA for 72 hours followed by treatment with test compound, each of which are set forth in Table 6, at either 4.15 ?M or 41.5 ?M. Data represents mean of either TNF? or interleukin IL-6 lowering after ConA stimulation?SD. 2-way ANOVA with Dunnet's multiple comparison test; * p<0.05 (represented as +); ** p<0.01 (represented as ++); *** p<0.001 **** (represented as +++); p<0.0001 (represented as ++++); N=3 replicates. Both TNF? and IL-6 are inflammatory cytokines for which the lowering would be expected to provide therapeutic benefit for a variety of diseases. For example, TNF? is implicated in the inflammation associated with neurodegenerative and inflammatory disorders (see, e.g., G. D. Kalliolias et al., TNF biology, pathogenic mechanisms and emerging therapeutic strategies, 12(1) NATURE REVIEWS RHEUMATOLOGY 49-62 (2016)) as is IL-17 (see, e.g., S. Kaur et al., A panoramic review of IL-6: Structure, pathophysiological roles and inhibitors, 28(5) BIOORG. MED. CHEM. 115327 (2020)).

(60) TABLE-US-00006 TABLE 6 Effect of Test Compounds on the Level of IL-6 and TNF? Lowering in ConA Stimulated Peripheral Blood Mononuclear Cells Molecular Concentration of test Weight compounds (?M) determined IL-6 TNF? # Test compound (daltons) 4.15 41.5 4.15 41.5 1 L-Arg-L-Lys-L-Asp-L-Val-L-Tyr (TP-5) 680.3 + +++ +++ ++++ (SEQ ID NO: 6) 2 D-Tyr-D-Val-D-Asp-D-Lys-D-Arg 680.3 ++ +++ ++ ++ (SEQ ID NO: 1) 3 D-Tyr-D-Val-D-Asp-D-Lys-D-Asp-NH.sub.2 638.2 ? +++ + ++ (SEQ ID NO: 7) 4 Ac-D-Tyr-D-Val-D-Asp-D-Lys-D-Arg- 721.3 + ++ ++ +++ NH.sub.2 (SEQ ID NO: 3) 5 D-Tyr-D-Val-D-Asp-D-Lys-D-Arg- 693.3 + ++ ? + NHMe (SEQ ID NO: 4) 6 Ac-D-Tyr-D-Val-D-Asp-D-Lys-D-Arg- 735.3 + ++++ ++++ ++++ NHMe (SEQ ID NO: 5)

Example 4TVALA? Effects Purinergic Signaling In In Vitro Stressed ALS-Derived Adipose Stem Cells

(61) Cells derived from ALS patient tissues exhibit hallmark metabolic defects that can be rescued when tested in vitro. See J-H. Hor et al., ALS motor neurons exhibit hallmark metabolic defects that are rescued by SIRT3 activation, 28 CELL DEATH & DIFFERENTIATION 1379-1397 (2021). Adipose stem cells (ASCs) derived from fat tissue harvested from an ALS patient were glycolytic showing increased proliferation and lipid accumulation in culture. An in vitro model of mitochondrial dysfunction was initiated with ALS patient derived ASCs grown in growth media. After cells were confluent, the growth media was treated with chemical stressors, such as ionomycin. Ionomycin induces cell death, apoptosis, peroxide formation, extracellular ATP, and peroxynitrites that are related to mitochondrial dysfunction. The media was exchanged and replaced with basic DMEM with 4 mM pyruvate and no additional growth factors. Test compounds were added in an effort to rescue the ionomycin treated cells. After recovery, 6 hours, the cells were measured for viability, apoptosis, peroxides, and peroxynitrites by SeaHorse analysis. The ASC patient derived cells are a good model to investigate dysregulated mitochondrial dysfunction in disease, these effects are not apparent in all ALS patient derived iPSC fibroblast cells (data not shown).

(62) Results of the analysis showed that ALS-ASC cells were hypermetabolic and retained lipids. The TVALA? treatment reduced peroxides and decreased extracellular ATP (eATP). See FIG. 5 (the line indicates a reduction of eATP with TVALA?). The eATP inhibition in this assay is expected to be pluribeneficial because purinergic signaling targets common responses in neurodegenerative disease, normalizing a dysregulated eATP may slow the progress of disease such as ALS). As stated previously, the complex cellular cross-talk occurring in ALS and the recognized function of extracellular nucleotides and adenosine in neuroglia communication, the ability of TVALA? to restoring purinome dynamics is beneficial in an ALS patient to slow the progression of disease as demonstrated in vitro.

Example 5Effect of TVALA? on Erastin-Induced Ferroptotic Cell Death

(63) Melanoma cell lines A-375, A-375 non-target, and A-375 (Prdx6 KO) cells were pre-incubated with TVALA?, washed, and treated with Erastin. Following treatment, the cells were evaluated for viability. Pretreatment with TVALA? did not rescue the cells against Erastin-induced cell death. See FIGS. 6A-6D. When the cells were pre-treated with TVALA?, washed, and incubated with TVALA? followed by treatment with Erastin, the cells were protected against Erastin-induced cell death. See FIGS. 7A-7D. When A-375 non target cells or A-375 knock out cells were used there was no effect of pre-treatment followed by TVALA? treatment as expected (data not shown). The TVALA? effect in the positive control assay, the A-375 cell line, demonstrates the ability to measure the effect of TVALA? in blocking Erastin-induced ferroptosis in additional cell lines such as cultured neurons.

Example 6D- but not L-Peptides have Significant Positive Effects

(64) ALS patient-derived ASCs showing hypermetabolism and lipid sequestration were stressed and treated with peptides (shown below, column 2) and effect of the peptide measured with SeaHorse analytics. The results show that the D- but not the L-peptides had a significant positive effect. TVALA? was superior to the other compounds.

(65) TABLE-US-00007 TABLE 7 Conc. with Test Range positive Compound Test compound (DMSO/H.sub.2O) effect p value 1 Arg-Lys-Asp-Val-Tyr (TP-5) 0.01-100 mM none (SEQ ID NO: 6) 2 Arg-Lys-Asp-Val-Tyr-NH.sub.2 5 mM none (SEQ ID NO: 8) 3 Ac-Arg-Lys-Asp-Val-Tyr-NH.sub.2 0.5 mM and none (SEQ ID NO: 9) 5 mM 4 Arg-Lys-Asp-Val-Tyr-NHMe 5 mM none (SEQ ID NO: 10) 5 Ac-Arg-Lys-Asp-Val-Tyr-NHMe 5 mM none (SEQ ID NO: 11) 6 D-Tyr-D-Val-D-Asp-D-Lys-D-Arg 5 mM 5 mM 0.0147 (SEQ ID NO: 1) 7 D-Tyr-D-Val-D-Asp-D-Lys-D-Arg-NH.sub.2 0.5 mM and 0.5 mM 0.0481 (SEQ ID NO: 2) 5 mM 8 Ac-D-Tyr-D-Val-D-Asp-D-Lys-D-Arg- 0.5 mM and 5 mM 0.0361 NH.sub.2 (SEQ ID NO: 3) 5 mM 9 D-Tyr-D-Val-D-Asp-D-Lys-D-Arg-NHMe 5 mM (SEQ ID NO: 4) 10 Ac-D-Tyr-D-Val-D-Asp-D-Lys-D-Arg- 5 mM NHMe (SEQ ID NO: 5)

Example 7TVALA? is Steroid Sparing

(66) TVALA? may be steroid sparing in diseases that involve innate immunity and autoimmune components that are traditionally treated with steroids. For example, age-related macular degeneration is treated with steroid anti-inflammatory agents. TVALA? is used as an adjunct to traditional therapies, decreasing or eliminating the need for steroids. Another example is for renal dialysis patients, for which TVALA? is used as adjunctive or as a replacement to anti-inflammatory therapies. Elamipretide Triacetate is a small, mitochondrially-targeted d-tetrapeptide that is not entirely different from TVALA?, as both have two basic residues (Arg and Lys) and both have a d-Arg. Elamipretide triacetate appears to reduce the production of the toxic reactive oxygen species and stabilize cardiolipin to moderate mitochondrial disease. In vitro comparisons of Elamipretide triacetate and TVALA? indicate TVALA? has more target receptors and is superior at immunomodulation that the tetrapeptide (data not shown).

Example 8TVALA? is Useful in Assays, Including in Discovering Novel Biomarkers

(67) Evaluating the effect of TVALA? in in vitro assays by incorporating patient derived cells and age/sex matched cells from clinically normal people as a control allow the evaluation of altered disease pathways. The identified factors that influence dysregulated pathways can reveal novel biomarkers. The lack of identifying biomarkers for neurodegenerative diseases to date point out that more dynamic and down-stream molecules need to be identified. In vitro assays incorporating patient cells allow a precision medicine approach. Using drugs such as TVALA? in assay systems identify areas of interest and allow stratification of patients.

Example 9TVALA? Modulates Exosomes

(68) Neurodegenerative diseases are closely related to brain function and the progression of the diseases are irreversible. Due to brain tissue being not easy to acquire, the study of the pathophysiology of neurodegenerative disorders has many limitations-lack of reliable early biomarkers and personalized treatment. At the same time, the blood-brain barrier (BBB) limits most of the drug molecules into the damaged areas of the brain, which makes a big drop in the effect of drug treatment. Exosomes, types of endogenous nanoscale vesicles, play a key role in cell signaling through the transmission of genetic information and proteins between cells. Because of the ability to cross the BBB, exosomes are expected to link peripheral changes to CNS events as potential biomarkers, and can even be used as a therapeutic carrier to deliver molecules specifically to CNS. Exosomes have a role in pathophysiology, diagnosis, prognosis, and treatment of some neurodegenerative diseases (including Alzheimer's Disease, Parkinson's Disease, Huntington's Disease, and Amyotrophic Lateral Sclerosis).

(69) Experiments show a similar profile of 32 growth factors and cytokines when culture media from adipose stem cells (ASC), dental pulp stem cells (DPSC), or mesenchymal stem cells (MSC) sources are evaluated. A similar but not identical profile is seen in ASCs derived from clinically normal patients and ALS derived ASCs. See FIG. 8 (normal patients) and FIG. 9 (ASC-ALS derived cells). Various growth factor formulations are tested and optimized. Expression of ASC VEGF-A, VEGF-D, VEGFR2, and VEGFR3 expression is measured. The VEGF pathway is an active area of investigation concerning hypermetabolism in ALS patients.

(70) TABLE-US-00008 ASC-CM DPSC-CM MSC-CM Growth Factors G-CSF ? ? ? VEGF ? ? ? EGF ? ? ? PDGF-BB ? ? ? b-NGF ? ? ? FGFb ? ? ? IGF-1 ? ? ? TGF-b ? ? ? PIGF-1 ? ? ? Cytokines TNFa ? ? ? IFN ? ? ? GM-CSF ? ? ? IL-1a ? ? ? IL-8 ? ? ? IP-10 ? ? ? Rantes ? ? ? IL-6 ? ? ? ? ? ? ? Resistin ? ? ? PAI-1 ? ? ? ? ? IL-12 ? ? ? IL-13 ? ? ? Eotaxin-3 ? ? ? SCF ? ? ? MCP-1 ? ? ? ? ? ? MIP-1a ? ? ? IL-2 ? ? ? IL-4 ? ? ? IL-10 ? ? ? Leptin ? ? ? Adipo ? ? ? IL-17a ? ? ? IL-1b ? ? ?

(71) Exosomes are membrane-bound extracellular vesicles. Exosomes can be diagnostic tools and cell signaling molecules. Exosomes can be a vehicle for drug delivery and therapy.

(72) TVALA? is incorporated into exosomes to exert beneficial effects in ALS. Adipose stem cell culture media is optimized for therapy in ALS. Exosomes from a commercial source are examined and compared to exosomes found in patients with neurodegenerative diseases. TVALA? rescues dysregulated mitochondrial energy metabolism in vitro, reduces extracellular peroxides in vitro (see FIG. 5), and normalizes VEGFR2 expression of the ALS patient derived cells. TVALA? is superior to TP-5 in this context. By incorporating TVALA? into exosomes and administering the exosome-spiked ASC-CM to patients, the therapy normalizes functional pathways that are diseased in neurodegenerative diseases that follow a final common pathway (such as AD, ALS, Huntington's, and Parkinson's).

CONCLUSIONS

(73) There are no ALS animal model systems that replicate the human phenotype. It is possible to use murine models to suggest target tissues to extrapolate to humans, however it is necessary to use in vitro assays from normal and ALS derived cells. The inventor's data shows using in vitro modeling that TVALA? affects cytokine pro-and anti-inflammatory levels as well as rescuing purinergic signaling in dysregulated cells. These responses are related to maintaining neuronal/glial tissue homeostasis.

Embodiments

(74) Embodiments of the invention include, but are not limited to, the following embodiment.

(75) Embodiment 1: A polypeptide comprising, in order from N-terminus to C-terminus,
R-X.sub.1-X.sub.2-X.sub.3-X.sub.4-X.sub.5-R wherein: R is optionally 1-5 additional ?-amino acids of either L-configuration or D-configuration; X.sub.1 is any amino acid of either L-configuration or D-configuration; X.sub.2 is any amino acid of either L-configuration or D-configuration; X.sub.3 is Asp or Glu of either L-configuration or D-configuration; X.sub.4 is any amino acid of either L-configuration or D-configuration; X.sub.5 is any amino acid of either L-configuration or D-configuration; and R is optionally 1-5 additional ?-amino acids of either L-configuration or D-configuration, with the proviso that at least three of X.sub.1, X.sub.2, X.sub.3, X.sub.4, and X.sub.5 are of the D-configuration, wherein the N-terminus is optionally modified by acetylation, and wherein the C-terminus is optionally modified by amidation and/or methylation.
Embodiment 2: The polypeptide of Embodiment 1, wherein said X.sub.1-X.sub.2-X.sub.3-X.sub.4-X.sub.5 is selected from the group consisting of: D-Tyr-D-Val-D-Asp-D-Lys-D-Arg (SEQ ID NO:1), D-Tyr-D-Val-D-Asp-D-Lys-D-Arg-NH.sub.2 (SEQ ID NO:2), Ac-D-Tyr-D-Val-D-Asp-D-Lys-D-Arg-NH.sub.2 (SEQ ID NO:3), D-Tyr-D-Val-D-Asp-D-Lys-D-Arg-NHMe (SEQ ID NO:4), Ac-D-Tyr-D-Val-D-Asp-D-Lys-D-Arg-NHMe (SEQ ID NO:5), and D-Tyr-D-Val-D-Asp-D-Lys-D-Asp-NH.sub.2 (SEQ ID NO:7).
Embodiment 3: The polypeptide of Embodiment 1, wherein R is H, X.sub.1 is D-Tyr, X.sub.2 is D-Val, X.sub.3 is D-Asp, X.sub.4 is D-Lys, X.sub.5 is D-Arg, and R is OH (SEQ ID NO:1).
Embodiment 4: A composition comprising a polypeptide according to any one of Embodiments 1-3 and a pharmaceutically-acceptable carrier.
Embodiment 5: A method for ameliorating a neurodegenerative disorder in a subject suffering from said neurodegenerative disorder comprising administering to said subject an effective amount of a polypeptide comprising, in order from N-terminus to C-terminus,
R-X.sub.1-X.sub.2-X.sub.3-X.sub.4-X.sub.5-R wherein: R is optionally 1-5 additional ?-amino acids of either L-configuration or D-configuration; X.sub.1 is any amino acid of either L-configuration or D-configuration; X.sub.2 is any amino acid of either L-configuration or D-configuration; X.sub.3 is Asp or Glu of either L-configuration or D-configuration; X.sub.4 is any amino acid of either L-configuration or D-configuration; X.sub.5 is any amino acid of either L-configuration or D-configuration; and R is optionally 1-5 additional ?-amino acids of either L-configuration or D-configuration, with the proviso that at least three of X.sub.1, X.sub.2, X.sub.3, X.sub.4, and X.sub.5 are of the D-configuration, wherein the N-terminus is optionally modified by acetylation, and wherein the C-terminus is optionally modified by amidation and/or methylation.
Embodiment 6: The method according to Embodiment 5, wherein said neurodegenerative disorder is selected from the group consisting of amyotrophic lateral sclerosis (ALS), Alzheimer's Disease (AD), dysregulated mitochondrial energy metabolism, impaired neuromuscular regulatory function, and Parkinson's Disease (PD).
Embodiment 7: The method according to Embodiment 5, wherein said X.sub.1-X.sub.2-X.sub.3-X.sub.4-X.sub.5 is selected from the group consisting of: D-Tyr-D-Val-D-Asp-D-Lys-D-Arg (SEQ ID NO:1), D-Tyr-D-Val-D-Asp-D-Lys-D-Arg-NH.sub.2 (SEQ ID NO:2), Ac-D-Tyr-D-Val-D-Asp-D-Lys-D-Arg-NH.sub.2 (SEQ ID NO:3), D-Tyr-D-Val-D-Asp-D-Lys-D-Arg-NHMe (SEQ ID NO:4), Ac-D-Tyr-D-Val-D-Asp-D-Lys-D-Arg-NHMe (SEQ ID NO:5), and D-Tyr-D-Val-D-Asp-D-Lys-D-Asp-NH.sub.2 (SEQ ID NO:7).
Embodiment 8: The method according to Embodiment 5, wherein R is H, X.sub.1 is D-Tyr, X.sub.2 is D-Val, X.sub.3 is D-Asp, X.sub.4 is D-Lys, X.sub.5 is D-Arg, and R is OH (SEQ ID NO:1).
Embodiment 9: The method according to Embodiment 5, wherein said polypeptide is administered intrathecally, intravenously, orally, and/or subcutaneously.
Embodiment 10: The method according to Embodiment 5, wherein said polypeptide is administered in an amount ranging from 0.1 mg/kg of said subject to 50 mg/kg of said subject.
Embodiment 11: The method according to Embodiment 5, wherein said polypeptide is administered in an amount ranging from 0.1 mg/kg of said subject to 10 mg/kg of said subject.
Embodiment 12: The method according to Embodiment 5, wherein said polypeptide is administered in an amount ranging from 0.1 mg/kg of said subject to 1 mg/kg of said subject.
Embodiment 13: The method according to Embodiment 5, wherein said polypeptide is lyophilized and reconstituted with an appropriate amount of diluent selected from the group consisting of distilled water and/or sodium chloride.
Embodiment 15: The method according to Embodiment 14, wherein said inflammatory disorder is selected from the group consisting of ankylosing spondylitis, chronic renal disease, Crohn's disease, dysregulated inflammatory response, hidradenitis suppurativa, inflammation, juvenile idiopathic arthritis, non-radiographic axial spondyloarthritis, non-infectious uveitis, neuroinflammation, plaque psoriasis, psoriasis, psoriatic arthritis, renal disease, rheumatoid arthritis, and ulcerative colitis.
Embodiment 16: The method according to Embodiment 14, wherein said X.sub.1-X.sub.2-X.sub.3-X.sub.4-X.sub.5 is selected from the group consisting of: D-Tyr-D-Val-D-Asp-D-Lys-D-Arg (SEQ ID NO:1), D-Tyr-D-Val-D-Asp-D-Lys-D-Arg-NH.sub.2 (SEQ ID NO:2), Ac-D-Tyr-D-Val-D-Asp-D-Lys-D-Arg-NH.sub.2 (SEQ ID NO:3), D-Tyr-D-Val-D-Asp-D-Lys-D-Arg-NHMe (SEQ ID NO:4), Ac-D-Tyr-D-Val-D-Asp-D-Lys-D-Arg-NHMe (SEQ ID NO:5), and D-Tyr-D-Val-D-Asp-D-Lys-D-Asp-NH.sub.2 (SEQ ID NO:7).
Embodiment 17: The method according to Embodiment 14, wherein R is H, X.sub.1 is D-Tyr, X.sub.2 is D-Val, X.sub.3 is D-Asp, X.sub.4 is D-Lys, X.sub.5 is D-Arg, and R is OH (SEQ ID NO:1).
Embodiment 18: The method according to Embodiment 14, wherein said polypeptide is administered intrathecally, intravenously, orally, and/or subcutaneously.
Embodiment 19: The method according to Embodiment 14, wherein said polypeptide is administered in an amount ranging from 0.1 mg/kg of said subject to 50 mg/kg of said subject.
Embodiment 20: The method according to Embodiment 14, wherein said polypeptide is administered in an amount ranging from 0.1 mg/kg of said subject to 10 mg/kg of said subject.
Embodiment 21: The method according to Embodiment 14, wherein said polypeptide is administered in an amount ranging from 0.1 mg/kg of said subject to 1 mg/kg of said subject.
Embodiment 22: The method according to Embodiment 14, wherein said polypeptide is lyophilized and reconstituted with an appropriate amount of diluent selected from the group consisting of distilled water and/or sodium chloride.